5-FU promotes HBV replication through oxidative stress-induced autophagy dysfunction

• Published in: Free radical biology & medicine Vol. 213; pp. 233 - 247
• Main Authors: Yang, Jing, Zheng, Luyan, Yang, Zhenggang, Wei, Zhiqiang, Shao, Jiajia, Zhang, Yina, Yao, Jiping, Li, Minwei, Wang, Xueyu, Zheng, Min
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.03.2024
• Subjects: Autophagic host defense; Chemotherapy; Oxidative stress; Viral reactivation; Autophagic host defense; CRP; Oxidative stress; BNIP3; Viral reactivation; HBeAg; TACE; HCC; SOD; MDA; PHH; Chemotherapy; SHBsAg; ROS; mTOR; HBcAg; HBV; LC3; 5-FU
• ISSN: 0891-5849; 1873-4596; 1873-4596
• DOI: 10.1016/j.freeradbiomed.2024.01.011

Hepatitis B virus (HBV) reactivation is a major problem that must be overcome during chemotherapy for HBV-related hepatocellular carcinoma (HCC). However, the mechanism underlying chemotherapy-associated HBV reactivation is still not fully understood, hindering the development of improved HBV-related HCC treatments. A meta-analysis was performed to assess the HBV reactivation risk during transcatheter arterial chemoembolization (TACE). To investigate the regulatory effects and mechanisms of 5-FU on HBV replication, an HBV mouse model was established by pAAV-HBV1.2 hydrodynamic injection followed by intraperitoneal 5-FU injection, and different in vitro models (HepG2.2.15 or Huh7 cells) were established. Realtime RT‒qPCR, western blotting, luciferase assays, and immunofluorescence were used to determine viral parameters. We also explored the underlying mechanisms by RNA-seq, oxidative stress evaluation and autophagy assessment. The pooled estimated rate of HBV reactivation in patients receiving TACE was 30.3 % (95 % CI, 23.1%–37.4 %). 5-FU, which is a chemotherapeutic agent commonly used in TACE, promoted HBV replication in vitro and in vivo. Mechanistically, 5-FU treatment obviously increased autophagosome formation, as shown by increased LC3-II levels. Additionally, 5-FU impaired autophagic degradation, as shown by marked p62 and mCherry-GFP-LC3 upregulation, ultimately promoting HBV replication and secretion. Autophagy inhibition by 3-methyladenine or chloroquine significantly altered 5-FU-induced HBV replication. Furthermore, 5-FU-induced autophagy and HBV replication were markedly attenuated with a reactive oxygen species (ROS) scavenger. Together, our results indicate that ROS-induced autophagosome formation and autophagic degradation play a critical role in 5-FU-induced HBV reactivation. [Display omitted] •Clinically TACE is associated with a higher failure rate of achieving HBV clearance.•5-FU delays clearance of HBV in vivo and promotes HBV replication in cell models.•Delayed HBV clearance is accompanied by increased levels of oxidative stress indicators.•5-FU activates autophagy initiation via ROS-BNIP3 to promote HBV replication.•5-FU blocks autophagosome degradation and promotes HBV replication.